RESUMEN
Insect prophenoloxidases (PPOs) are important immunity proteins for defending against the invading pathogens and parasites. As a Type-â ¢ copper-containing proteins, unlike Homo sapiens tyrosinases, the insect PPOs and most bacterial tyrosinases contain no signal peptides for unknown reason, however they can still be released. To this end, we fused different signal peptides to Drosophila melanogaster PPOs for in vitro and in vivo expression, respectively. We demonstrate that an artificial signal peptide can help PPO secretion in vitro. The secreted PPO appeared larger than wild-type PPO on molecular weight sizes due to glycosylation when expressed in S2 cells. Two asparagine residues for potential glycosylation in PPO1 were identified when a signal peptide was fused. After purification, the glycosylated PPO1 lost zymogen activity. When PPO1 containing a signal peptide was over-expressed in Drosophila larvae, the glycosylation and secretion of PPO1 was detected in vivo. Unlike insect PPO, human tyrosinase needs a signal peptide for protein expression and maintaining enzyme activity. An artificial signal peptide fused to bacterial tyrosinase had no influence on the protein expression and enzyme activity. These Type-â ¢ copper-containing proteins from different organisms may evolve to perform their specific functions. Intriguingly, our study revealed that the addition of calcium inhibits PPO secretion from the transiently cultured larval hindguts in vitro, indicating that the calcium concentration may regulate PPO secretion. Taken together, insect PPOs can maintain enzyme activities without any signal peptide.
Asunto(s)
Catecol Oxidasa , Drosophila melanogaster , Precursores Enzimáticos , Señales de Clasificación de Proteína , Animales , Catecol Oxidasa/metabolismo , Precursores Enzimáticos/metabolismo , Drosophila melanogaster/inmunología , Drosophila melanogaster/metabolismo , Glicosilación , Humanos , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Larva/metabolismo , Precursores de Proteínas/metabolismo , Monofenol Monooxigenasa/metabolismo , Línea Celular , Proteínas de Insectos/metabolismo , Proteínas de Insectos/genética , Calcio/metabolismoRESUMEN
Maintaining a definite and stable pool of dividing stem cells plays an important role in organ development. This process requires an appropriate progression of mitosis for proper spindle orientation and polarity to ensure the ability of stem cells to proliferate and differentiate correctly. Polo-like kinases (Plks)/Polo are the highly conserved serine/threonine kinases involved in the initiation of mitosis as well as in the progression of the cell cycle. Although numerous studies have investigated the mitotic defects upon loss of Plks/Polo in cells, little is known about the in vivo consequences of stem cells with abnormal Polo activity in the context of tissue and organism development. The current study aimed to investigate this question using the Drosophila intestine, an organ dynamically maintained by the intestinal stem cells (ISCs). The results indicated that the polo depletion caused a reduction in the gut size due to a gradual decrease in the number of functional ISCs. Interestingly, the polo-deficient ISCs showed an extended G2/M phase and aneuploidy and were subsequently eliminated by premature differentiation into enterocytes (ECs). In contrast, the constitutively active Polo (poloT182D) suppressed ISC proliferation, induced abnormal accumulation of ß-tubulin in cells, and drove ISC loss via apoptosis. Therefore, Polo activity should be properly maintained for optimal stem cell function. Further analysis suggested that polo was a direct target gene of Sox21a, a Sox transcription factor that critically regulates stem cell activity. Together, this study provided a novel perspective on the correlation between the progression of mitosis and the ISC function in Drosophila.
Asunto(s)
Proteínas de Drosophila , Drosophila , Animales , Ciclo Celular/genética , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Drosophila/genética , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Intestinos , Mitosis/genética , Células Madre/metabolismoRESUMEN
After harvest, cassava (Manihot esculenta Crantz) storage roots undergo rapid postharvest physiological deterioration, producing blue-brown discoloration in the vasculature due to the production of polyphenolics (mainly quinones and coumarins) by enzymes such as polyphenol oxidase (PPO). Here, we report the application of hen egg-white lysozyme (HEWL), a natural PPO inhibitor, in transgenic cassava to repress the symptoms of postharvest physiological deterioration. The HEWL-expressing transgenic plants had lower levels of the two main cassava coumarins tested, scopoletin and scopolin, compared with wild type. HEWL-expressing cassava also showed increased tolerance of oxidative stress. Overall, the lysozyme-PPO system proved to be functional in plants for repressing PPO-mediated commercial product browning.
Asunto(s)
Manihot , Manihot/genética , Muramidasa/genética , Raíces de Plantas , Plantas Modificadas Genéticamente , EscopoletinaRESUMEN
Prophenoloxidase (PPO), an important immunity protein in insects, is mainly produced by hemocytes and released into the hemolymph upon cell lysis. In addition, PPO can also be produced by epidermal cells in the foregut to detoxify the toxic plant secondary metabolites and in the hindgut to kill pathogens through PPO-induced melanization. Previously, we noticed a pair of tubes extended from the larval hindgut became melanized upon staining in dopamine dissolved in 30% ethanol. However, the structure and function of these tubes are largely unknown. In this study, we performed staining of the tubes and the neighboring Malpighian tubule for further confirmation. Eventually, we detected PPO inside epidermal cells of the tubes, and called them as PPO-positive tubes. We observed that the PPO-positive tubes are physically derived from the hindgut but strongly adhere to the Malpighian tubule. Inside the PPO-positive tubes, there is an acellular peritrophic membrane to protect the epidermal cells. Furthermore, the PPO-positive tubes act like a doorkeeper to firstly detoxify the metabolite wastes collected by the Malpighian tubule from the hemolymph.
Asunto(s)
Lepidópteros , Túbulos de Malpighi , Animales , Catecol Oxidasa/metabolismo , Precursores Enzimáticos/metabolismo , Túbulos de Malpighi/metabolismoRESUMEN
Entomopathogenic fungi Beauveria bassiana can infect many species of insects and is used as a biological pesticide world-wide. Before reaching the hemocoel, B. bassiana has to penetrate the integument which is composed of a thick chitin layer and epidermal cells. Some chitinase, protease and lipase secreted by B. bassiana are probably involved in the fungal penetration of the integument. While microscopic proof is needed, it is difficult to locate the precise infection sites following the traditional method of immersion infection. Consequently, we developed a new method to inoculate conidia solution into a single fixed-site on the back of one segment. This fixed-site infection method is pathogenic but it is also dose dependent. Using the fixed-site infection protocol, it is also very convenient to track hyphae inside the cuticle layer by light and transmission electron microscopy. The fact that few hyphae were detected inside the chitin layer after fixed-site infection with mutant ΔBPS8, a protease secreted during fungi germination, indicates that this method is suitable for screening genes involved in penetrating the integument in large scale. We also found that melanization occurs before new hyphae penetrate the chitin layer. Most importantly, we discovered that fungal infection can induce epidermal cell proliferation through DNA duplication and cell division, which is essential for the host to defend against fungal infection. Taken together the fixed-site infection method may be helpful to determine the mechanism of fungal and host interaction in the integument so as to effectively exert fungal biological virulence.
Asunto(s)
Beauveria/fisiología , Bombyx/inmunología , Quitina/metabolismo , Epidermis/metabolismo , Micosis/inmunología , Animales , Proliferación Celular , Quitinasas/metabolismo , Epidermis/patología , Interacciones Microbiota-Huesped , Hifa , Proteínas de Insectos/metabolismo , Lipasa/metabolismo , Microscopía Electrónica de Transmisión , Mutación/genética , Péptido Hidrolasas/metabolismo , Control de Plagas , Esporas Fúngicas , VirulenciaRESUMEN
Insects depend on the innate immune response for defense against a wide array of pathogens. Central to Drosophila immunity are antimicrobial peptides (AMPs), released into circulation when pathogens trigger either of the two widely studied signal pathways, Toll or IMD. The Toll pathway responds to infection by Gram-positive bacteria and fungi while the IMD pathway is activated by Gram-negative bacteria. During activation of the IMD pathway, the NF-κB-like transcription factor Relish is phosphorylated and then cleaved, which is crucial for IMD-dependent AMP gene induction. Here we show that loss-of-function mutants of the unconventional histone variant H2Av upregulate IMD-dependent AMP gene induction in germ-free Drosophila larvae and adults. After careful dissection of the IMD pathway, we found that Relish has an epistatic relationship with H2Av. In the H2Av mutant larvae, SUMOylation is down-regulated, suggesting a possible role of SUMOylation in the immune phenotype. Eventually we demonstrated that Relish is mostly SUMOylated on amino acid K823. Loss of the potential SUMOylation site leads to significant auto-activation of Relish in vivo. Further work indicated that H2Av regulates Relish SUMOylation after physically interacting with Su(var)2-10, the E3 component of the SUMOylation pathway. Biochemical analysis suggested that SUMOylation of Relish prevents its cleavage and activation. Our findings suggest a new mechanism by which H2Av can negatively regulate, and thus prevent spontaneous activation of IMD-dependent AMP production, through facilitating SUMOylation of the NF-κB like transcription factor Relish.
Asunto(s)
Proteínas de Drosophila/metabolismo , Histonas/metabolismo , Inmunidad Innata/genética , Factores de Transcripción/metabolismo , Animales , Péptidos Catiónicos Antimicrobianos/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/inmunología , Proteínas de Drosophila/fisiología , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Expresión Génica/genética , Regulación de la Expresión Génica/genética , Histonas/fisiología , FN-kappa B/genética , Transducción de Señal/genética , Sumoilación/genética , Receptores Toll-Like , Factores de Transcripción/genéticaRESUMEN
Gut microbiome profoundly affects many aspects of host physiology and behaviors. Here we report that gut microbiome modulates aggressive behaviors in Drosophila. We found that germ-free males showed substantial decrease in inter-male aggression, which could be rescued by microbial re-colonization. These germ-free males are not as competitive as wild-type males for mating with females, although they displayed regular levels of locomotor and courtship behaviors. We further found that Drosophila microbiome interacted with diet during a critical developmental period for the proper expression of octopamine and manifestation of aggression in adult males. These findings provide insights into how gut microbiome modulates specific host behaviors through interaction with diet during development.
Asunto(s)
Agresión/fisiología , Drosophila melanogaster/fisiología , Microbioma Gastrointestinal/fisiología , Octopamina/metabolismo , Conducta Sexual Animal/fisiología , Animales , Bacterias/clasificación , Bacterias/genética , Encéfalo/citología , Encéfalo/metabolismo , Encéfalo/fisiología , Drosophila melanogaster/metabolismo , Drosophila melanogaster/microbiología , Femenino , Microbioma Gastrointestinal/genética , Masculino , Actividad Motora/fisiología , Neuronas/metabolismo , Neuronas/fisiología , ARN Ribosómico 16S/genética , Transducción de Señal/fisiología , Organismos Libres de Patógenos EspecíficosRESUMEN
Bt protein, produced by Bacillus thuringiensis, can bind receptors to destroy the physiological functions of the insect midgut. It is unknown whether Bt can also target the hindgut and influence its defense against fecal bacteria. Here we show that Crystal protein 1Ab (Cry1Ab), a Bt protein, was detected in the larval hindgut contents of Bombyx mori after ingestion of this toxin protein. The number of fecal bacteria that can be inhibited by the hindgut prophenoloxidase-induced melanization was significantly enhanced after oral ingestion of Cry1Ab. Although the hindgut contents became brown, the activity of hindgut phenoloxidase was decreased. LC-MS/MS analysis of the hindgut lumen contents revealed that many new proteins including several proteases were newly secreted. The enhanced secretion of proteases cleaved prophenoloxidase to decrease its activity, including the corresponding activity to inhibit the fecal bacteria. In addition, after ingestion of Cry1Ab, the pylorus (between the midgut and hindgut) could not autonomously contract due to the physical detachment of the acellular cuticle-like membrane from the epidermal cells, which prevented the movement of food from the midgut to the hindgut. Some cells in the cryptonephry of the hindgut became swollen and degraded, possibly due to the presence of Cry1Ab in the hindgut. These findings demonstrate that the inhibition of feces bacteria by the hindgut prophenoloxidase-induced melanization is out of control after Cry1Ab ingestion.
Asunto(s)
Toxinas de Bacillus thuringiensis/metabolismo , Bacillus thuringiensis/fisiología , Bombyx/inmunología , Sistema Digestivo/metabolismo , Endotoxinas/metabolismo , Infecciones por Bacterias Grampositivas/inmunología , Proteínas Hemolisinas/metabolismo , Animales , Catecol Oxidasa/metabolismo , Cromatografía Liquida , Sistema Digestivo/patología , Ingestión de Alimentos , Precursores Enzimáticos/metabolismo , Proteínas de Insectos/metabolismo , Larva , Espectrometría de Masas en TándemRESUMEN
Insects produce many peptide hormones that play important roles in regulating growth, development, immunity, homeostasis, stress, and other processes to maintain normal life. As part of the digestive system, the insect midgut is also affected by hormones secreted from the prothoracic gland, corpus allatum, and various neuronal cells; these hormones regulate the secretion and activity of insects' digestive enzymes and change their feeding behaviors. In addition, the insect midgut produces certain hormones when it recognizes various components or pathogenic bacteria in ingested foods; concurrently, the hormones regulate other tissues and organs. In addition, intestinal symbiotic bacteria can produce hormones that influence insect signaling pathways to promote host growth and development; this interaction is the result of long-term evolution. In this review, the types, functions, and mechanisms of hormones working on the insect midgut, as well as hormones produced therein, are reviewed for future reference in biological pest control.
RESUMEN
A diverse family of metalloproteases (MPs) is distributed in eukaryotes. However, the functions of MPs are still understudied. We report that seven MPs belonging to the M35 family are encoded in the genome of the insect pathogenic fungus Metarhizium robertsii. By gene deletions and insect bioassays, we found that one of the M35-family MPs, i.e. MrM35-4, is required for fungal virulence against insect hosts. MrM35-4 is a secretable enzyme and shows a proteolytic activity implicated in facilitating fungal penetration of insect cuticles. After gene rescue and overexpression, insect bioassays indicated that MrM35-4 contributes to inhibiting insect cuticular and hemocyte melanization activities. Enzymatic cleavage assays revealed that the recombinant prophenoloxidases PPO1 and PPO2 of Drosophila melanogaster could be clipped by MrM35-4 in a manner differing from a serine protease that can activate PPO activities. In addition, it was found that MrM35-4 is involved in suppressing antifungal gene expression in insects. Consistent with the evident apoptogenic effect of MrM35-4 on host cells, we found that the PPO mutant flies differentially succumbed to the infections of the wild-type and mutant strains of M. robertsii. Thus, MrM35-4 plays a multifaceted role beyond targeting PPOs during fungus-insect interactions, which represents a previously unsuspected strategy employed by Metarhizium to outmaneuver insect immune defenses.
Asunto(s)
Catecol Oxidasa/metabolismo , Precursores Enzimáticos/metabolismo , Insectos/microbiología , Metaloproteasas/genética , Metarhizium/genética , Metarhizium/metabolismo , Animales , Apoptosis , Drosophila melanogaster/microbiología , Femenino , Proteínas Fúngicas/genética , Eliminación de Gen , Regulación Fúngica de la Expresión Génica , Interacciones Huésped-Patógeno , Larva/microbiología , Metaloproteasas/metabolismo , Metarhizium/enzimología , Virulencia/genéticaRESUMEN
Entomopathogenic fungi are promising bio-pesticides. To facilitate infection, fungi recruit multiple virulence factors and deploy different molecular strategies to evade host immunity. Fungal ribotoxins are extracellular secreted ribonucleases (RNases) with ribotoxic cytotoxicity and insecticidal activity. However, it remains unclear whether they have further biological functions. Here we show that the entomopathogenic fungus Beauveria bassiana ribotoxin (Rib) contributes to fungal virulence by inhibiting insect host immunity. Gene deletion of Rib (ΔRib) resulted in attenuated fungal virulence during infection. Pathogenesis analysis demonstrated that Rib mainly inhibits insect immunity through modulating the reactive oxygen species (ROS) response, suppressing antimicrobial peptides (AMPs) production and retarding hyphae penetration from insect cuticles. To further confirm this immunosuppressive function, recombinant ribotoxin (rRib) protein was purified and co-injected with living or heat-killed bacteria, bacteria-derived peptidoglycan (PGN) and lipopolysaccharide (LPS) separately, which also significantly inhibited the AMPs production in Drosophila fat bodies. Furthermore, co-injection of rRib with Escherichia coli or Staphylococcus aureus significantly enhanced bacterial pathogenicity and facilitated infection. In addition, rRib injection resulted in a global inhibition of protein expression in different tissues of Drosophila adults. This work identified B. bassiana ribotoxin as a key virulence factor that inhibits insect immunity.
Asunto(s)
Beauveria/fisiología , Infecciones por Escherichia coli/inmunología , Escherichia coli/fisiología , Micotoxinas/genética , Staphylococcus aureus/fisiología , Animales , Células Cultivadas , Eliminación de Gen , Hifa , Evasión Inmune , Inmunidad Innata , Micotoxinas/metabolismo , Proteínas Citotóxicas Formadoras de Poros , VirulenciaRESUMEN
Aluminum (Al) is an important environmental metal factor that can be potentially associated with pathological changes leading to neurotoxicity. The silkworm, Bombyx mori, is an important economic insect and has also been used as a model organism in various research areas. However, the toxicity of Al on silkworm physiology has not been reported. Here, we comprehensively investigate the toxic effects of Al on the silkworm, focusing on its effects on viability and development, superoxide dismutase (SOD) activity, and the expression of presenilin and cAMP response element-binding protein (CREB) in BmE cells and silkworm larvae. BmE cell viability decreased after treatment with aluminum chloride (AlCl3 ) in both dose- and time-dependent manners. When AlCl3 solution was injected into newly hatched fifth instar larvae, both larval weight gain and survival rate were significantly decreased in a manner correlating with AlCl3 dose and developmental stage. Furthermore, when BmE cells and silkworm larvae were exposed to AlCl3 , SOD activity decreased significantly relative to the control group, whereas presenilin expression increased more than twofold. Additionally, CREB and phosphorylated CREB (p-CREB) expression in the heads of fifth instar larvae decreased by 28.0% and 50.0%, respectively. These results indicate that Al inhibits the growth and development of silkworms in vitro and in vivo, altering SOD activity and the expressions of presenilin, CREB, and p-CREB. Our data suggest that B. mori can serve as a model animal for studying Al-induced neurotoxicity or neurodegeneration.
Asunto(s)
Compuestos de Aluminio/toxicidad , Aluminio/toxicidad , Bombyx/efectos de los fármacos , Cloruros/toxicidad , Contaminantes Ambientales/toxicidad , Proteínas de Insectos/genética , Neurotoxinas/toxicidad , Cloruro de Aluminio , Animales , Peso Corporal/efectos de los fármacos , Bombyx/enzimología , Bombyx/genética , Bombyx/crecimiento & desarrollo , Línea Celular , Supervivencia Celular/efectos de los fármacos , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Proteínas de Insectos/metabolismo , Larva/efectos de los fármacos , Larva/enzimología , Larva/genética , Longevidad/efectos de los fármacos , Presenilinas/genética , Presenilinas/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismoRESUMEN
Insect prophenoloxidase (PPO) induces melanization around pathogens. Before melanization, PPO is cleaved into phenoloxidase (PO) by serine proteases. Insect PPO can also be activated by exogenous proteases secreted by pathogens as well as by other compounds, such as ethanol and cetylpyridinium chloride (CPC). However, the effect of these activators on the activity of PO is unclear. In this study, the insect endogenous serine protease AMM1, α-chymotrypsin, and ethanol were used to activate recombinant Drosophila PPO1 (rPPO1), and the PO activity differed depending on the activator applied. The PO-induced intermediates during melanization also varied markedly in their numbers and abundances. Therefore, this study indicates that the mechanism of PPO activation influences PO activity. It also suggests that PO-induced different intermediates may affect the antibacterial activity during melanization due to their toxicity.
Asunto(s)
Catecol Oxidasa/genética , Drosophila melanogaster/genética , Precursores Enzimáticos/genética , Levodopa/metabolismo , Monofenol Monooxigenasa/genética , Animales , Catecol Oxidasa/metabolismo , Drosophila melanogaster/metabolismo , Precursores Enzimáticos/metabolismo , Monofenol Monooxigenasa/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
Signal peptidase complexes (SPCs) are conserved from bacteria to human beings, and are typically composed of four to five subunits. There are four genes encoding SPC proteins in the red flour beetle, Tribolium castaneum. To understand their importance to insect development, double-stranded RNA for each SPC gene was injected into red flour beetles at the early larval and adult stages. Knockdown of all four signal peptidase genes was lethal to larvae. Moreover, larvae had difficulty with old cuticle ecdysis. Knockdown of TcSPC12 alone did not affect pupal or adult development. When TcSPC12, TcSPC18, and TcSPC25 were knocked down in larvae, the melanization of hemocytes and midguts was observed. When knocked down in larvae and adults, TcSPC18 induced severe cell apoptosis in midguts, and the adult midgut lost the ability to maintain crypts after knockdown of TcSPC18, indicating its importance to midgut cell proliferation and differentiation. Knockdown of TcSPC22 or TcSPC25 also resulted in many apoptotic cells in the midguts. However, TcSPC12 appeared to be unimportant for midgut development. We conclude that TcSPC18 is essential for maintaining the adult midgut crypts.
Asunto(s)
Proteínas de la Membrana/metabolismo , Serina Endopeptidasas/metabolismo , Tribolium/enzimología , Animales , Femenino , Tracto Gastrointestinal/enzimología , Hemocitos/metabolismo , Proteínas de Insectos/metabolismo , Melaninas/metabolismo , Interferencia de ARN , Tribolium/crecimiento & desarrolloRESUMEN
Skin immunity protects animals from airborne pathogen infection. Unlike mammals, arthropods, including insects, undergo periodic ecdysis to grow and develop. Newly molted insects emerge with unsclerotized thin cuticles but successfully escape pathogenic infections during the post-molt period. Here we show that prophenoloxidases (PPOs) in molting fluids remain bioactive on the integument and impede fungal infection after ecdysis. We found that the purified plasma PPOs or recombinant PPOs could effectively bind to fungal spores (conidia) by targeting the cell wall components chitin and ß-1,3-glucan. Pretreatment of the spores of the fungal pathogen Beauveria bassiana with PPOs increased spore hydrophilicity and reduced spore adhesion activity, resulting in a significant decrease in virulence as compared with mock infection. We also identified a spore-secreted protease BPS8, a member of peptidase S8 family of protease that degrade PPOs at high levels to benefit fungal infection, but which at lower doses activate PPOs to inhibit spore germination after melanization. These data indicate that insects have evolved a distinct strategy of ex vivo immunity to survive pathogen infections after ecdysis using PPOs in molting fluids retained on the underdeveloped and tender integument of newly molted insects for protection against airborne fungal infection.
RESUMEN
Integuments are the first line to protect insects from physical damage and pathogenic infection. In lepidopteran insects, they undergo distinct morphology changes such as scale formation during metamorphosis. However, we know little about integument development and scale formation during this stage. Here, we use the silkworm, Bombyx mori, as a model and show that stem cells in the integument of each segment, but not intersegmental membrane, divide into two scale precursor cells during the spinning stage. In young pupae, the scale precursor cell divides again. One of the daughter cells becomes a mature scale-secreting cell that undergoes several rounds of DNA duplication and the other daughter cell undergoes apoptosis later on. This scale precursor cell division is crucial to the development and differentiation of scale-secreting cells because scale production can be blocked after treatment with the cell division inhibitor paclitaxel. Subsequently, the growth of scale-secreting cells is under the control of 20-hydroxyecdysone but not juvenile hormone since injection of 20-hydroxyecdysone inhibited scale formation. Further work demonstrated that 20-hydroxyecdysone injection inhibits DNA duplication in scale-secreting cells while the expression of scale-forming gene ASH1 was down-regulated by BR-C Z2. Therefore, this research demonstrates that the scale cells of the silkworm develops through stem cell division prior to pupation and then another wave of cell division differentiates these cells into scale secreting cells soon after entrance into the pupal stage. Additionally, DNA duplication and scale production in the scale-secreting cells were found to be under the regulation of 20-hydroxyecdysone.
Asunto(s)
Diferenciación Celular/fisiología , Ecdisona/metabolismo , Regulación del Desarrollo de la Expresión Génica , Lepidópteros/metabolismo , Animales , Bombyx/metabolismo , Células Epiteliales/metabolismo , Hormonas Juveniles/metabolismo , Metamorfosis Biológica/fisiología , Pupa/metabolismoRESUMEN
BACKGROUND: Insects can be models for understanding human intestinal infection and pathology. Molting, a special period during which the old insect cuticle is shed and a new one is produced, is crucial for insect development. Holometabolous insects may experience several larva-to-larva moltings to become larger, a pupal molt and adult eclosion to become adults. During the larval molts, they stop feeding and become quiescent. Although the molting larvae become quiescent, it is not known if changes in microbiome, physiology, development and immunity of midguts occur. RESULTS: Transcriptome analysis indicated that functions such as metabolism, digestion, and transport may become reduced due to the downregulated expression of many associated genes. During the molting stage, midguts harbor less microflora and DNA synthesis decreases. Both ecdysone and juvenile hormone in the larval midgut likely degrade after entering the larva-to-larva molting stage. However, at 12 h after ecdysis, the feeding larvae of 5th instars that were injected with 20-hydroxyecdysone entered a molting-like stage, during which changes in midgut morphology, DNA synthesis, gene expression, and microflora exhibited the same patterns as observed in the actual molting state. CONCLUSION: This study is important for understanding insect midgut physiology, development and immunity during a special development stage when no food is ingested. Although the molting larva becomes immobile and quiescent, we demonstrate that numerous changes occur in midgut morphology, physiology, metabolism and microbiome during this period.
Asunto(s)
Bombyx/crecimiento & desarrollo , Bombyx/genética , Sistema Digestivo/metabolismo , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Muda/genética , Animales , Transporte Biológico , Análisis por Conglomerados , Metabolismo Energético , Microbioma Gastrointestinal , Larva , Estadios del Ciclo de Vida , TranscriptomaRESUMEN
Lepidopteran insects constitute one of the largest fractions of animals on earth, but are considered pests in their relationship with man. Key to the success of this order of insects is its ability to digest food and absorb nutrition, which takes place in the midgut. Because environmental microorganisms can easily enter Lepidopteran guts during feeding, the innate immune response guards against pathogenic bacteria, virus and microsporidia that can be devoured with food. Gut immune responses are complicated by both resident gut microbiota and the surrounding peritrophic membrane and are distinct from immune responses in the body cavity, which depend on the function of the fat body and hemocytes. Due to their relevance to agricultural production, studies of Lepidopteran insect midgut and immunity are receiving more attention, and here we summarize gut structures and functions, and discuss how these confer immunity against different microorganisms. It is expected that increased knowledge of Lepidopteran gut immunity may be utilized for pest biological control in the future.
Asunto(s)
Infecciones Bacterianas/inmunología , Células Caliciformes/fisiología , Inmunidad Mucosa , Intestinos/inmunología , Lepidópteros/inmunología , Microsporidiosis/inmunología , Virosis/inmunología , Animales , Intestinos/microbiología , Microbiota , Control de PlagasRESUMEN
Invertebrate animals have the capacity of repairing wounds in the skin and gut via different mechanisms. Gastrointestinal perforation, a hole in the human gastrointestinal system, is a serious condition, and surgery is necessary to repair the perforation to prevent an abdominal abscess or sepsis. Here we report the repair of gastrointestinal perforation made by a needle-puncture wound in the silkworm larval midgut. Following insect gut perforation, only a weak immune response was observed because the growth of Escherichia coli alone was partially inhibited by plasma collected at 6 h after needle puncture of the larval midgut. However, circulating hemocytes did aggregate over the needle-puncture wound to form a scab. While, cell division and apoptosis were not observed at the wound site, the needle puncture significantly enhanced DNA duplication in cells surrounding the wound, which was essential to repair the midgut perforation. Due to the repair capacity and limited immune response caused by needle puncture to the midgut, this approach was successfully used for the injection of small compounds (ethanol in this study) into the insect midgut. Consequently, this needle-puncture wounding of the insect gut can be developed for screening compounds for use as gut chemotherapeutics in the future.